Complete Chloroplast Genome Sequence Structure and Phylogenetic Analysis of Brassica juncea var. multiceps (Brassicaceae)

Brassica juncea var. multiceps (Xuelihong), a variety of B. juncea (L.) Czern., holds considerable nutritional and economic value. However, its complete chloroplast genome and the evolutionary relationships within Brassicaceae remain poorly characterized. Using Illumina NovaSeq 6000 high-throughput sequencing, we assembled and annotated the full chloroplast genome sequence of B. juncea var. multiceps. The genome is 153,483 bp in length, with 36.36% GC content, and encodes 132 genes. Codon usage analysis identified leucine (Leu) as the dominant amino acid. Thirty-one codons had relative synonymous codon usage (RSCU; a metric for codon preference) values greater than one, with 93.55% of these preferred codons ending in A/U. We detected 37 dispersed repeats (14 forward, 18 palindromic, 3 reverse, and 2 complementary) and 315 simple sequence repeats (SSRs), with mononucleotide SSRs dominating (72.70%). Analysis of the Ka/Ks ratio, a measure of selection pressure (where values greater than one indicate positive selection), indicated that ycf1, ycf2, and nadhF genes may have undergone positive selection. The nucleotide diversity analysis revealed five hypervariable hotspot-genomic regions with high mutation rates, which are critical for phylogenetic studies. Phylogenetic analysis of 26 Brassicaceae species revealed that B. juncea var. multiceps is closely related to B. juncea. Notably, this is the first complete chloroplast genome of B. juncea var. multiceps, with unique hypervariable regions not reported in other B. juncea varieties. These findings clarify evolutionary relationships in Brassicaceae, provide molecular markers for the genetic breeding of B. juncea var. multiceps, and enhance our understanding of chloroplast genome adaptation in Brassica.